Cyan-green-fluorescent protein

A bright cyan-green fluorescent protein was isolated from Clavu-laria coral [86]. Since one of the intermediates displayed fast bleaching, a screen for more photostable variants was performed. The optimized monomeric variant was named teal fluorescent protein 1 (mTFPl). It has an excitation and emission maximum at 462 and 492 nm, respectively, so this protein is spectrally located in between CFP and GFP. With an extinction coefficient of 64,000 M 1 cm-1 and a quantum yield of 0.85 mTFPl is a very bright fluorescent protein. 

DOTMA E. coli E EBV ECFP ECV EGFP ELISA EYFP FACS FdG FH2 FH4 FK506 FLP propane-aminium-trifluoracetate 7V-[2,3-(dioleyloxy) propyl]-/V,/V,/V-trimethyl ammonium chloride Escherichia coli erythromycine operon/repressor Epstein-Barr virus enhanced cyan fluorescence protein extracellular viral particles enhanced green fluorescence protein enzyme-linked immunosorbent assay enhanced yellow fluorescence protein fluorescence-activated cell sorter fluorescein di- 3-D-galactopyranoside dihydrofolate tetrahydrofolate human immunophilins native recombinase isolated from the 2pm plasmid from Saccharomyces cerevisiae... 

Volkmer A, Subramaniam V, Birch DJS, Jovin TM. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins. Biophys. J. 2000 78 1589-1598. Subramaniam V, Hanley QS, Clayton AHA, Jovin TM. Photophysics of green and red fluorescent proteins implications for quantitative microscopy. Methods Enzymol. 2003 360 178-201. Rizzo MA, Springer GH, Granada B, Fdston DW. An improved cyan fluorescent protein variant useful for FRET. Nature Biotechnol. 2004 22 445-449. 

Further studies using green fluorescent protein or cyan fluorescent protein tagged receptors showed that a1B/a1D-AR heterodimerization appeared to completely control the surface expression and functional coupling of a1D-ARs on the plasma membrane (76). Coexpression of a1B, but not a1A-, ARs resulted in almost exclusively surface localization of the normally intracellular a1D-ARs, consistent with the specificity observed in previous coimmunoprecipitation studies. Further studies showed that the hydrophobic core of the a1B-AR is the major structural determinant of this interaction, and that G protein coupling was not required (76). These studies suggest that subtype-specific heterodimerization of ARs may control surface expression, and that these observations maybe relevant to many other class I G protein-coupled receptors, for which the functional consequences of this phenomenon are still poorly understood. 

ABSTRACT A number of fluorescent proteins have been discovered in marine organisms with the green-fluorescent protein (GFP) from Aequorea victoria representing the first member of this family being isolated and well characterized. These polypeptides show marked differences in their spectral properties. Today, blue-, yellow-, cyan- and red-light emitting proteins are known in addition to GFP. 

EBFP Enhanced Blue Fluorescent Protein ECFP Enhanced Cyan Fluorescent Protein EGFP Enhanced Green Fluorescent Protein EYFP Enhanced Yellow Fluorescent Protein. 

Living cell microscopy has imdergone taken a major leap forward in the last ten years, thanks to the introduction of natural fluorescent probes such as green fluorescent protein (GFP) [2-4] and its derivatives (cyan FP, yellow FP, etc.). These advances, combined with increases in the sensitivity of high-res-olution detectors, have made it possible to observe hving cells for a long... 

Fig. 2 List of some of the most widely employed fluorochromes in FM with indication of their peaks of excitation and emission. On the right are indicated the main lines of emission of a mercury lamp (ML) and four common laser sources. According to the wavelength of emitted light, lasers can be indicated as UV lasers (350 nm), violet lasers (407 nm), Wue lasers (488 nm), and ref/lasers (635 nm). 7-A4D7-aminoactinomycin 0, APC allophycocyanin, CFP cyan fluorescent protein, Em emission (nm). Ex excitation (nm), EYFP enhanced yellow fluorescent protein, FAf fluorescence microscopy, F/ Pfluorescent reporter protein, 6FF green fluorescent protein, IMF immunofluorescence, mito mitochondrion/mitochondrial, ML mercury lamp, RFP red fluorescent protein, variant of YFP, KFFyellowfluorescentprotein...

Proteins that exhibit fluorescence in their native form were extensively studied and utilized in a variety of apphcations. One important example is green fluorescent protein (GFP) of Aequorea victoria and its mutants (BFP, CFP, YFP), which exhibit blue, cyan, and yeUow emissions, respectively. In addition, a novel fluorescent protein, termed DsRed, was recently cloned and characterized. These proteins are unique in having fluorophores formed from the natural amino acid side chains via cyclization. A major application for them is the creation and expression of fluorescent fusion proteins. Such fusion constructs were used in a variety of apphcations involving in vitro and in vivo spatial and temporal fluorescence... 

The first described member of the cyan fluorescent proteins (CFPs) resulted from a rationally designed chromophore mutation of Aequorea GFP. Heim and co-worker replaced Tyr66 with Trp and found the peak wavelength for excitation and emission of this GFP derivative (GFP-Y66W) to be shifted to 436 and 476 nm, respectively [52], Because of this blue-green/cyan light emission the protein was called cyan fluorescent protein or CFP. 

CFP Cyan fluorescent protein. CFP is a derivative of the green fluores-... 

Various mutations of the GFP gene have resulted in fluorescent proteins colored blue, cyan, yellow, and red, in addition to green (Lippincott-Schwartz and Patterson, 2003). Other fluorescent proteins from reef coral have been discovered and have been optimized for expression in mammalian cells (BD Biosciences, 2004). 

FITC+FAM (fluorescein derivatives). ECFP, EGFP, EYFP (enhanced—cyan, green and yellow fluorescent protein, respectively, Clontech Laboratories, Inc., USA). Bodipy is a trademark of Invitrogen Ltd., UK. Attoxxxis a trademark of ATID-TEC GmbH, Gennany. A A S, and A J), A are the one- and two-photon absorption and emission maximum, respectively. QY is the fluorescence quantum yield, SS the Stokes shift, e the molar extinction coefficient and Tf the fluorescence lifetime. Further information regarding two-photon measurements can be found In Chapter 3. 

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